Retardation of metalloproteinase incidental to HIV and/or AIDS

ABSTRACT

The instant invention provides a product of manufacture for retarding the biochemical formation of metalloproteinase, including gelatinase, elastase, collaginase, and the like, within the tissues of the body of a patient who has been inflicted with the HIV virus and/or the HIV virus which has advance to the AIDS virus, through the administration of an effective amount of a non-antimicrobial/non-antibiotic/non-antibacterial, chemically modified tetracycline (CMT) analog, its salts, cojugates and/or derivatives, and combinations thereof.

BACKGROUND OF THE INVENTION

This is an application for the Reissue of U.S. Pat. No. 6,063,775,published May 16, 2000.

The instant invention broadly relates to retardation of the biochemicalformation of materials incidental to HIV and/or AIDS, which promotetheadvancement of HIV to AIDS; which promote the advancement of AIDS andother opportunistic diseases to which a patient could inevitablysuccumb. Specifically, the invention retards the biochemical formationof species of metalloproteinase which are formed as a product ofconditions such HIV and/or AIDS. Still more specifically, the inventionprovides for the administration of an effective amount of a drug whichretards the biochemical formation of metalloproteinase species includingcoliagenase and gelatinase, incidental to the conditions of HIV and/orAIDS. Still even more specifically, the invention provides for theadministration of an effective amount of a chemically modifiedtetracycline (CMT) analog which retards the biochemical formation ofmetalloproteinase species including gelatinase, collagenase andelastase, incidental to the conditions of HIV and/or AIDS.

Tetracyclines are useful as broad spectrum antibiotics because they havethe ability to retard protein synthesis in a wide variety of bacteria.As disclosed in the above-identified pending patent applications, it hasalso been discovered that tetracyclines, antibiotic tetracyclines andnon-antibiotic tetracyclines, have the ability to retardcollagen-destructive enzymes, such as collagenase, responsible for thebreak-down of connective tissue in a number of diseases, such asperiodontal disease, corneal ulcers and rheumatoid arthritis.

The use of tetracycline antibiotics, while effective, may lead toundesirable side effects. For example, the long term administration ofantibiotic tetracyclines may reduce or eliminate healthy flora, such asintestinal flora, and may lead to the production of antibiotic resistantorganisms or the overgrowth of yeast and fungi.

Tetracycline may be a chemically modified tetracycline (CMT) or anytetracycline administered to a mammal in a dose that is effectivelynon-antimicrobial in the mammal. Preferably, the tetracycline ischemically modified so as to reduce its antimicrobial properties.Methods for reducing the antimicrobial properties of a tetracycline aredisclosed in “The Chemistry of the Tetracyclines”, Chapter 6, Mitscher,(1978), at page 211. As pointed out by Mitscher, modification apositions 1, 2, 3, 4, 10 and 12a lead to loss of bioactivity. The use ofsuch modified tetracyclines is preferred in the present invention, sincethey can be used to higher levels than antimicrobial tetracyclines withfewer side effects.

The tetracycline molecule is amenable to substantial modificationwithout losing its antibiotic properties. Examples of modifications thatmay and may not be made to the basic tetracycline structure have beenreviewed by Mitscher in the Chemistry of Tetracyclines, Chapter 6.According to Mitscher, the substituents at positions 5-9 of thetetracycline ring, may be modified without complete loss of antibioticproperties. Changes to the basic ring system or replacement of thesubstituents at positions 1-4 and 10-12, however, generally lead tosynthetic tetracyclines having substantially less or effectively noantibacterial activity. For example, 4-dedimethylaminotetracycline iscommonly considered to be a non-antibacterial tetracycline.

U.S. Pat. No. 5,532,227 to Golub, et al teaches a method for treatingmammals suffering from excessive extracellular protein glycosylationwhich is associated with diabetes, scleroderma and progeria byadministering to the mammal a tetracycline which effectively inhibitsexcessive protein glycosylation.

Inhibition of metalloproteinase activity with various species ofnon-antimicrobial tetracycline, is well known in the prior art:

U.S. Pat. No. 5,321,017 to Golub, et al teaches a method for treatingmammals suffering from rheumatoid arthritis and other tissue-destructive(chronic inflammatory or other) conditions associated with excessmetalloproteinase activity which comprises: administering to the mammalan amount of a tetracycline that is effectively anti-metalloproteinase,but that is not effectively antimicrobial, and an amount ofnon-steroidal anti-inflammatory agent which, when combined with theeffectively anti-metalloproteinase amount of tetracycline, results in asignificant reduction of tissue destruction and/or bone loss.

U.S. Pat. No. 5,045,538 to Schneider, et al teaches a method fortreating mammals suffering from skeletal muscle wasting and/orintracellular protein degradation of skeletal muscle systems byadministering to the mammal an amount of tetracycline which results in asignificant reduction of the muscle wasting and protein degradation. Inaddition, there is also disclosed a method of increasing the proteincontent of skeletal muscle systems of mammals by administration oftetracyclines. The tetracyclines useful in the above methods are bothantimicrobial and non-antimicrobial. In a preferred embodiment, themethod of treatment utilizes a non-antimicrobial tetracycline such asdedimethylaminotetracycline.

U.S. Pat. No. 5,324,634 to Zucker teaches diagnostic agents and methodsfor detecting the presence of metastatic activity in biological samplessuch as plasma. The agent and method preferably immunologically detectmatrix metalloproteinases in complexed form with endogenous inhibitorsof MMP's A kit for detecting the metalloproteinases is also disclosed.

U.S. Pat. No. 5,260,059 to Acott, et al relates to a method of treatingocular disease by modulating cellular secretion of a family of matrixmetalloproteinases and their inhibitor. Specifically, differentialstimulation of secretion of interstitial collagenase, gelatinase or typeIV collaginase, stromelysin or proteoglycanase, and their tissueglycoprotein inhibitor is employed to treat open-angle glaucoma, retinaldegeneration and detachment, ocular neovascularization and diabeticretinopathy.

U.S. Pat. No. 5,595,885 to Stetler-Stevenson, et al teaches an isolatedprotein of 21,600 Da which binds to both latent and activated type IVcollagenase with high affinity at 1:1 molar stoichiometry, therebyabolishing enzyme activity. The protein is purified by affinitychromatography on solid phase metalloproteinase, or solid phasemetalloproteinase substrates which bind the enzyme-inhibitor complex.The complete primary structure of this protein (initially calledCSC-21K), as determined by sequencing overlapping peptides spanning theentire protein, reveals homology with a protein called TIMP, TissueInhibitor of Metalloproteinases. In addition, a cDNA for this novelinhibitor, now designated TIMP-2, was cloned from a melanoma cell andits sequence was compared with that of human TIMP-1. Northern blots ofmelanoma cell mRNA showed two distinct transcripts of 0.9 kb and 3.5 kbwhich are down-regulated by transforming growth factor-beta, and areunchanged by phorbol ester treatment. The inhibitor of the presentinvention may be used for treatment of pathologic conditions resultingfrom inappropriate degradation of extracellular matrix molecules bymatrix metalloproteinases, such as metastatic neoplasia, myocardialinfarction, and arthritis. Therapeutic treatments using this inhibitormay include formulations for inhalation and inclusion complexes adaptedfor buccal or sublingual administration, or administration of arecombinant DNA molecule which expresses a DNA segment that encodes thematrix metalloproteinase inhibitor of this invention.

U.S. Pat. No. 5,308,839 to Golub, et al teaches a method for treatingmammals suffering from rheumatoid arthritis, other tissue-destructiveconditions, and chronic inflammatory or other conditions associated withexcess metalloproteinase activity which comprises: administering to themammal an amount of a tetracycline that is effectivelyanti-metalloproteinase, but that is not effectively antimicrobial, andan amount of a non-steroidal anti-inflammatory agent which, whencombined with the effectively anti-metalloproteinase amount oftetracycline, results in a significant reduction of tissue destructionand/or bone loss.

U.S. Pat. No. 5,532,227 to Golub, et al teaches a method for treatingmammals suffering from excessive extracellular protein glycosylationwhich is associated with diabetes, scleroderma and progeria byadministering to the mammal a tetracycline which effectively inhibitsexcessive protein clycosylation.

U.S. Pat. No. 5,308,839 to Golub, et al teaches a method for treatingmammals suffering from rheumatoid arthritis, other tissue-destructiveconditions, and chronic inflammatory or other conditions associated withexcess metalloproteinase activity comprising administering to the mammalan amount of a tetracycline that is effectively anti-metalloproteinase,but that is not effectively antimicrobial, and an amount of anon-steroidal anti-inflammatory agent which, when combined with theeffectively anti-metalloproteinase amount of tetracycline, results in asignificant reduction of tissue destruction and/or bone loss.

U.S. Pat. No. 5,223,248 to McNamara et al teaches a method of inhibitingplaque formation on mammalian tooth surfaces. The method includescontacting the tooth surfaces with an effective amount of anon-antibacterial tetracycline. In a preferred embodiment, suchtetracyclines are included in various oral hygiene products such asdentifrices, lozenges, chewing gums and the like to contact the toothsurfaces and prevent plaque accumulation thereon.

U.S. Pat. No. 5,045,538 in Schneider, et al teaches a method fortreating mammals suffering from skeletal muscle wasting and/orintercellular protein degradation of skeletal muscle systems byadministering to the mammal an amount of tetracycline which results in asignificant reduction of the muscle wasting and protein degradation. Inaddition, there is also disclosed a method of increasing the proteincontent of skeletal muscle systems of mammals by administration oftetracyclines. The tetracyclines useful in the above methods are bothantimicrobial and non-antimicrobial. In a preferred embodiment, themethod of treatment utilizes a non-antimicrobial tetracycline such asdedimethylaminotetracycline (CMT).

Generally, tetracyclines, as has now been discovered, whether possessingantimicrobial or antibiotic activity or not, all possessanti-collagen-destructive enzyme activity or anti-collagenase activity.They are known to inhibit the activity of collagen destructive enzymessuch as mammalian collagenase, macrophage elastase and bacterialcollagenase; Golub et al., J. Periodont. Res. 20, 12-23 (1985), Golub etal., Crit. Revs. Oral Biol. Med. 2, 297-332 (1991). Collagen is a majorcomponent of connective tissue matrices such as those in bone, synovium,eye, skin, tendons and gingiva. Collagenase, which is naturally producedby only a few types of bacteria and in a number of tissues and cells inmammals, degrades collagen.

This anti-collagenase activity appears to be attributable to the uniquestructure of tetracyclines, i.e. the special four carbocyclic ringstructure which is characteristic of and possessed by the tetracyclines.

As an observation, it is believed that the carbonyl moieties in thecarbocyclic ring nucleus of the tetracycline are important to theanti-collagenolytic activity of these compounds because they chelate themetal ions calcium and zinc. This is an important property since thecollagenolytic enzymes mentioned are metal dependent.

It is known that antimicrobial and non-antimicrobial tetracyclines canbind to metal ions such as calcium.

Tetracyclines are also known retarders of collagen destructive enzymessuch as mammalian collagenase, a calcium dependentzinc-metalloproteinase. Collagen is a major component of connectivetissue matrices such as those in the bone, synovium, eye, skin, tendonsand gingiva but not tooth surface enamel.

In the case of mammalian collagenase, degradation of collagen is anatural part of the normal growth-degradation-regeneration process thatoccurs in connective tissue. The production of collagenase, however, maybecome excessive. Such excessive collagenase production often results inthe pathologic and debilitating destruction of connective tissue.

It is well known that metalloproteinases which include collogenases areproduced as byproducts of HIV and AIDS.

It is known that HIV-infected monocytes form highly invasive network onbasement membrane matrix and secrete high levels of 92-kdmetalloproteinase (MMP-9), an enzyme that degrades basement membraneproteins. In a study, using matrigel as a model basement membranesystem, it was demonstrated that treatment of human immunodeficiencyvirus HIV-infected monocytes with interferon-gamma at 50 U/ml inhibitedthe ability of infected monocytes to form an invasive network onmatrigel and their invasion through the matrigel matrix. These effectswere associated with a significant reduction in the levels of MMP-9produced by HIV-infected monocytes treated with interferon-gamma 1 dayprior to infection with HIV as compared with that of untreatedHIV-infected monocytes. Monocytes treated with interferon-gamma 1 dayafter HIV infection showed the presence of integrated HIV sequences;however, the levels of MMP-9 were substantially lower than thoseproduced by monocytes inoculated with live HIV, heat-inactivated HIV, oreven the control uninfected monocytes. Exposure of monocytes toheat-inactivated HIV did not result in increased invasiveness or highMMP-9 production, suggesting that regulation of metalloproteinase bymonocytes was independent of CD4-gp 120 interactions and required activevirus infection. Furthermore, addition of interferon-gamma to monocyteson day 10 after infection inhibited MMP-9 production by more thanthreefold with no significant reduction of virus replication. It wasconcluded that these results indicate that the mechanism ofinterferon-gamma-induced down-regulation of MMP-9 levels and reducedmonocyte invasiveness may be mediated by a mechanism independent ofantiviral activity of IFN-gamma in monocytes. Down-regulation of MMP-9in HIV-infected monocytes by interferon-gamma may play an important rolein the control of HIV pathogenesis.

It has further been reported that monocytes are susceptible to HIVinfection and to activation by a regulatory gene product of the HIVgenome, HIV-Tat. Recently, it was demonstrated that treatment withHIV-Tat up-regulates monocyte adhesion to the endothelium and increasesmetalloproteinase production in the present study, the ability of theHIV-Tat protein to alter the migratory and invasive behavior ofmonocytes were examined. Monocytes pre-treated for 24 hours with 10ng/ml HIV-Tat exhibited enhanced migratory behavior compared withuntreated monocytes in chemotaxis assays, both in the absence of achemoattractant as well as in response to FMLP in addition, HIV-Tatitself induced the migration of both untreated and HIV-Tat pretreatedmonocytes. Checkerboard analysis showed that monocytes migrated inresponse to an HIV-Tat concentration gradient, thus confirming thechemotactic characteristics of the HIV-Tat protein. Pretreatment ofmonocytes with 10 ng/ml HIV-Tat for 24 h also increased their ability toinvade reconstituted extracellular membrane (Matrigel)-coated filters by5-fold in the absence of chemoattractant. The presence of FMLP orHIV-Tat further enhanced invasion by both untreated andHIV-Tat-pretreated monocytes by more than 10-fold. Monocyte invasion waspartially inhibited by the inclusion of anti-beta integrin Ab or tissueinhibitor of metalloproteinase (TIMP). Thus, for the first time,evidence was provided that HIV-Tat can enhance the chemotactic andinvasive behaviors of monocytes and propose an active role for HIV-Tatin the recruitment of monocytes into extravascular tissues, a processwhich may contribute to the destruction of tissues and cellulararchitecture often seen in patients with acquired immunodeficiencysyndrome.

It was further demonstrated HIV infection of monocytes resulted intwofold elevation of adhesion molecule LFA-1 (both alpha L/CD11a andbeta 2/CD18 subunits) and LFA-3 (CD58), with no apparent increase inLFA-2 (CD2) or various beta 1-integrins. Homotypic aggregation ofmonocytes was evident 2 hours after exposure to virus and was inhibitedby mAbs to both the alpha L- and beta 2-subunits of LFA-1. HIV-infectedmonocytes also showed a marked increase in adherence to brain capillaryendothelial cell monolayers derived from brain, lung, and skin. Thisadherence was inhibited by mAb to either LFA-1 subunit and by mAb to thecounter-receptor intercellular adhesion molecule-1. Cocultivation ofHIV-infected monocytes with endothelial cells increased permeability ofendothelial cell monolayers to 125I albumin in transwell assay systems.The increased endothelial permeability induced by HIV-infected monocyteswas associated with a substantial disruption of the endothelial cellmonolayer. Morphologic disruption was not a direct toxic effect onendothelial cells, but appeared to be secondary to changes inendothelial cell-cell to cell-matrix interactions. Northern blotanalysis showed increased expression of gelatinase B (92-kDagelatinase), tissue inhibitor of metalloproteinase TIMP-1, and TIMP-2 inthe-infected monocytes. Consistent with these Northern analyses,secretion of gelatinase activity in culture fluids of HIV-infectedmonocytes was also increased and was dependent on the staage of virusreplication. Incubation of HIV-infected monocytes with the proteinaseinhibitors TIMP-1 and TIMP-2 inhibited the increased permeability ofendothelial cell monolayers to 125I albumin. These results suggestpossible mechanisms for extravasation of HIV-infected monocytes throughvascular endothelium into tissue in early stages of HIV disease.

SUMMARY OF THE INVENTION

The instant invention in large part solves the problems of the prior andfulfills a long felt need by providing a drug in a dosage sufficient foreffectively retarding the biochemical formation of metalloproteinasewithin the bodies of patients inflicted with HIV and/or AIDS viruses.

Here are the more important features of the invention as broadlyoutlined, in order that the detailed description that follows may bebetter understood; and in order for the present contribution to the artmay be better appreciated. There are additional features of theinvention that will be described hereinafter and which form the subjectmatter of the appended claims. Those of ordinary skill in the art willappreciate that the conception upon which this disclosure is based mayreadily be utilized as a basis for the designing of other structures,methods and systems for carrying out the several purposes of the instantinvention. It is important, therefore, that the claims be regarded asincluding such equivalent constructions insofar as they do not departfrom the spirit and scope of the instant invention.

Further, the purpose of the instant abstract is to enable the U.S.Patent and Trademark office and the public generally, and especially thescientists, engineers and practitioners in the art who are not familiarwith parent or legal terms or phraseology, to determine quickly from acursory inspection of it, the technical disclosure of the patentapplication. The abstract is neither intended to define the invention ofthe instant patent application, which is measured by the claims, nor isit intended in any manner to be limiting as to the scope of the instantinvention.

The instant invention in large part solves the problems of the prior andfulfills a long felt need by providing a treatment for a patient withHIV and/or AIDS.

The instant invention provides a drug in a dosage sufficient fortreating a patient who has been inflicted with the HIV and/or AIDSviruses, with an effective dosage of anon-antimicrobial/non-antibiotic/non-antibacterial, drug in the body ofthe patient.

The instant invention provides a drug to a dosage sufficient fortreating a patient who has been inflicted with the HIV and/or AIDSviruses, with an effective dosage of anon-antimicrobial/non-antibiotic/non-antibacterial, chemically modifiedtetracycline (CMT) analog in the body of the patient.

The instant invention provides a drug in a dosage sufficient fortreating a patient who has been inflicted with the HIV and/or AIDSviruses, with an effective dosage of anon-antimicrobial/non-antibiotic/non-antibacterial, chemically modifiedtetracycline (CMT) sufficient to retard the biochemical formation ofmetalloproteinase within the body of the patient.

The instant invention provides a drug in a dosage sufficient fortreating a patient who is inflicted with the HIV and/or AIDS viruses,with an effective dosage of anon-antimicrobial/non-antibiotic/non-antibacterial, chemically modifiedtetracycline (CMT) analog effective to retard the biochemical formationof metalloproteinase within the body of the patient.

The instant invention provides a drug in a dosage sufficient fortreating a patient who is inflicted with the HIV and/or AIDS viruses,with an effective dosage of a chemically modified tetracycline (CMT)analog sufficient to retard the biochemical formation of gelatinase,elastase, collaginase, and the like, within the tissues of the body ofthe patient.

The instant invention provides a drug to a dosage sufficient fortreating a patient who is inflicted with the HIV and/or AIDS viruses,with an effective dosage of anon-antimicrobial/non-antibiotic/non-antibacterial, chemically modifiedtetracycline (CMT) analog sufficient to retard the biochemical formationof gelatinase, elastase, collaginase, and the like, within the tissuesof the body of the patient.

The instant invention provides a drug in a dosage sufficient fortreating a patient who is inflicted with the HIV and/or AIDS viruses,with an effective dosage of anon-antimicrobial/non-antibiotic/non-antibacterial, chemically modifiedtetracycline (CMT) analog sufficient to retard the biochemical formationof gelatinase, elastase, collaginase, and the like, within the tissuesof the body of the patient.

Other objects, features, and advantages of the instant invention, in itsdetails of construction and arrangement of parts, will be seen from theabove, from the following description of the preferred embodiment whenconsidered in light of the appended claims.

DETAILED DESCRIPTION OF THE INVENTION

The instant invention provides a drug in a dosage sufficient forretarding endrogenous metalloproteinase and at least its species ofelastase, collagenase and gelatinase, characterized by that occurring inthe body of a patient who has become inflicted with the HIV virus and/ortreating a patient who has become infected with the HIV virus which hasprogressed to AIDS. The instant invention contemplates administering tothe patient an effective dosage of an analog of chemically modifiedtetracycline (CMT) selected from the group consisting of:4-dedimethylaminotetracycline, 4-dedimethylamino-5-oxytetracycline,4-dedimethylamino-7-chlortetracycline,4-hydroxy-4-dedimethylaminotetracycline, 5a,6-anhydro-4hydroxy-4-dedimethylaminotetracycline,6alpha-deoxy-5-hydroxy-4-dedimethylaminotetracycline,6-demethyl-6-deoxy-4-dedimethylaminotetracycline,4-dedimethyaminno-11-hydroxy-12a-deoxytetracycine,12a-deoxy-4-deoxy-4-dedimethylaminotetracycline,6alpha-deoxy-5-hydroxy-4-dedimethylaminotetracycline,12a,4a-anhydro-4-dedimethylaminotetracycline, minocycline-CMT,7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,

6a-benzylthiomethylenetetracycline, the 2-nitrilo analogs oftetracycline (tetracyclinonitrile), the mono-N-alkylated amide oftetracycline; 6fluoro-6-demethyltetracycline, 11a-chloroletracycline,tetracycline pyrazole, 12a-deoxytetracycline and its derivatives,4-dedimethylamino-5oxytetracycline, 5a,6-anhydro-4-hydroxy-4-dedimethylaminotetracycline,12a,4a-anhydro-4-dedimethylaminotetracycline, tetracyclinonitrile,7-chloro-4-dedimethylaminotetracycline,12a-deoxy-4-deoxy-4-dedimethylaminotetracycline,4-dedimethylamino-7chlorotetracycline,4-dedimethylamino-7-dimethylamninoletracycline, the 2-nitrilo analogs oftetracycline, 4-dedimethylamino-12a-deoxytetracycline and itsderivatives, tetracyclines altered at the 2 carbon position to produce anitrile, 4-de-dimethylamino-7-chlorotetracycline,6-alpha-deoxy-5-hydroxy-4-dedimethylaminotetracycline,tetracyclinotrile, 6-alpha-benzylthiomethylenetetracycline, the2-nitrilo analog of tetracycline, 11 alpha-chlorotetracycline,7-chlorotetracycline, 5-hydroxytetracycline,6-demethyl-7-chlorotetracycline,6-demethyl,-6-deoxy-5-hydroxy-6-methylenetetracycline,6-alpha-benzylthiomethylenetetracycline, a nitrite analog oftetracycline, a mono-N-alkylated amide of tetracycline,11-alpha-chlorotetracycline, 2-acetyl-8-hydroxy-1-tetracycline,6-demethyl-6-deoxytetracycline,6-demethyl-6-deoxy-5-hydroxy-6-methylenetetracycline,2-acetyl-8-hydroxyl-1-tetracycline,4-hydroxy-4dedimethylaminotetracycline,5a,6-anhydro-4-hydroxy-4dedimethylaminotetracycline,6-demethyl-6-deoxy-4-dedimethylaminotetracycline,6-deoxy-6-demethyl-4-dedimethylaminotetracycline,6a-deoxy-5-hydroxy-4-dedimethylaminotetracycline, Tetracyclines alteredat the 2-carbon position to produce a nitrile, pyrazole derivative oftetracycline, 7-chloro-6-demethyl-4-dedimethylaminotetracycline, 11alpha-chlortetracycline, 4-dedimethylamino-7-chlortetracycline,4-de(dimethylamino)-tetracycline, 4-de(dimethylamino)-5-oxytetracycline,4-de(dimethylamino)-7-chlortetracycline, 11 alpha a-chlortetracycline,7-chloro-6-demethyl-4-dedimethylaminotetracycline,6-o-deoxy-5-hydroxy-4-dedimethylaminotetracycline,6-alpha-obenzylthiomethylenetetracycline,4-de(dimethylamino-5-oxytetracycline,4-de(dimethylamino)-7-chlorotetracycline,4-hydroxy-4-dedimethylaminotetracycline,6-alpha-deoxy-5-hydroxy-4-dedimethylamino-tetracycline,4-de(dimethylamino) tetracycline, 4-de(dimethylamino)-7chlorotetracycline, 7-chloro-6-demethyl-4-dedimethylamino-tetracycline,dedimethylaminotetracycline, a 6-alpha-benzylthiomethylene tetracycline,6-alpha benzylthiomethylene tetracycline, an 11-alpha-chlortetracycline,6-demethyl-6-deoxy-5-hydroxy-6-methylenetetracycline, a 6alpha-benzylthiomethylene tetracycline, and a 6 Fluorodemethyltetracycline, their salts, cojugates and/or derivatives, andcombinations thereof.

The analog of the instant invention may be administered orally, orsystemically, or by way of an injection or intravenously. It may also beadministered topically in the form of an ointment, or other storabletopical delivery system, i.e., applying it with a highly adsorptivematerial such as DMSO.

Dosages of the analog can range in amounts of from about 0.1 mg/kg/dayto about 50 mg/kg/day, more preferably in amounts of from about 10.0mg/kg/day to about 30 mg/kg/day; and most preferably amounts of about20.0 mg/kg/day to about 25.0 mg/kg/day.

The preferred pharmaceutical composition for use in the presentinvention comprises a combination of the chemically modifiedtetracycline (CMT) and the anti-inflammatory agent in a suitablepharmaceutical carrier. The means of delivery of the pharmaceuticalcarrier with active may be in the form of a capsule, compressed tablet,pill, solution or suspension suitable for oral administration to amammal. Other means of delivery include a gel for topical applicationfor corneal ulcers, periodontal disease, etc. It is contemplated thatcarriers be included which are suitable for administration orally,topically, by injection into a joint, and by other selected means.

The non-steroidal anti-inflammatory agent may be selected from thevarious classes of such compounds. Such classes include, for example,salicylates such as acetylsalicyclic acid and diflunisal, acetic acidssuch as indomethacin, sulindac, tolmetin, diclofenac, and etodolac;propionic acids such as flurbiprofen, naproxen, and ketoprofen;fenamates such as meclofenamate; and oxicams such as piroxicam.

The preferred non-steroidal anti-inflammatory agents includeflurbiprofen, piroxicam, tolmetin sodium, ibuprofen, naproxen andindomethacin. The preferred non-steroidal anti-inflammatory agentsinclude flurbiprofen, piroxicam, tolmetin sodium, ibuprofen, naproxen,indomethacin and tenidap. Tenidap (CP-66,248-2) is available from PfizerCentral Research (Groton, Conn.).

The non-steroidal anti-inflammatory agent may be selected from thevarious classes of such compounds. Such classes include, for example,salicylates such as acetylsalicyclic acid and diflunisal; acetic acidssuch as indomethacin, sulindac, tolmetin, diclofenac, and etodolac;propionic acids such as flurbiprofen, naproxen, indomethacin, tolmetinsodium and ketoprofen; fenamates such as meclofenamate; oxicams such aspiroxicam; and oxindoles such as tenidap.

The amount of the non-steroidal anti-inflammatory agent is an amountwhich, when combined with the effectively anti-collagenase amount oftetracycline, results in a significant reduction of bone loss in mammalssuffering from tissue-destructive conditions associated with excessmetalloproteinase activity. The amount depends on the particularanti-inflammatory agent used, the mammal to which the composition isadministered, and the amount of the tetracycline in the composition.Some typical doses for routine human use include, for example, 20 mg/dayfor piroxicam, 150 mg/day for indomethacin, 1600-1800 mg/day forlolmetin, 1000 mg/day for naproxen, and 3200 mg/day for ibuprofen.

For example, a suitable amount of a CMT such as 4-dedimethylaminotetracycline is 15 mg/kg. A suitable mount of anti-inflammatory agent incombination with 30 mg/kg of a CMT such as 4-dedimethylaminotetracycline would be, for example, 1-8 mg/kg flurbiprofen, 0.3 mg/kgpiroxicam and 40 mg/kg ibuprofen. As a guideline for providing theproper amount of anti-inflammatory agents for implementing the presentinvention, a rule of thumb is to administer an amount which is 20% to80% of the conventional anti-inflammatory dose for treating arthritis.Thus, the dosage could be from as small as 10 mg/person/day forpiroxicam, to as great as 3200 mg/person/day for ibuprofen. In anyevent, the practitioner is guided by skill and knowledge in the fieldand the present invention includes without limitation dosages which arceffective to achieve the described phenomenon. For example, thenon-steroidal anti-inflammatory agent my be administered in an amount offrom about 0.3 mg/kg per day to about 3,500 mg per person per day.

Thus, without undue experimentation, one of ordinary skill in the artcan readily vary the dosage, form and/or method of administration, withrespect to a particularly selected CMT of the instant invention, so asto provide a product of manufacture for the effective treatment for apatient inflicted with HIV and/or AIDS.

Although the invention preferably contemplates treatment of the tissuesof the body of a patient inflicted with the HIV and/or AIDS viruses, itis equally applicable to the treatment of any disease which causes thebiochemical formation of metalloproteinase and any of its species,elastase, gelatinase, collagenase, and the like, within any tissue inthe body of a patient.

The term “analog” and its variants as broadly used herein, is meant toinclude other related species such as homologs of tetracycline.

The terms “non-antimicrobial,” “non-antibiotic,” and“non-antibacterial,” are meant to have equivalent definitions as usedherein.

The term “biochemical” as used herein is intended to refer tobiotechnology.

Although the invention has been described with reference to certainpreferred embodiments, it will be appreciated that many variations andmodifications may be made within the scope of the broad principles ofthe invention. Hence, it is intended that the preferred embodiments andall of such variations and modifications be included within the scopeand spirit of the invention, as defined by the following claims.

I claim:
 1. A method for healing HIV viral infections, or AIDScomprising administering to patients in need of such treatment ananti-retroviral amount of non-antimicrobial, non-antibiotic,non-antibacterial chemically modified tetracycline (CMT) analogs.
 2. Themethod of claim 1, wherein the chemically modified tetracycline (CMT) isselected from the group consisting of: 4-dedimethylaliiinotetracycline,4-dedimtntnhylamino-oxytetracycline,4-dedimethylamino-7-chlortetracycline,4-hydroxy-4-dedimethylaminotetracycline,5a,6-anhydro-4-hydroxy-4-dedimethylaminotetracycline,6-alpha-dcoxy-5-hydroxy-4-dedimethylaminotetracycline,6-dernethyl-6-deoxy-4-dedimelhylaminotetracycline,4-dedimethylamino-11-hydroxy-12a-deoxytetracyclins,12a-deoxy-4-deoxy-4-dedimethylaminotetracycline,6-alpha-deoxy-5-hydroxy-4-dedimethylaminodoxycycline,12a,4a-anhydro-4-dedimethylaminotetracycline, minocycline-CMT,7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,6a-benzylthiomethylenetetracycline, the 2-nitrilo analogs oftetracycline (tetracyclinonitrile), the mono-N-alkylated amide oftetracycline, 6-fluoro-6-demethyltetracycline, 11a-chlortetracycline,tetracycline pyrazole, 12a-deoxytetracycline,4-dedimethylamino-5-oxytetracycline,5a,6-anhydro-4-hydroxy-4-dedimethylaminotetracycline,12a,4a-anhydro-4-dedimethylaminotetracycline, tetracyclonitrile,7-chloro-4-dedimethylaminotetracycline,12a-deoxy-4-deoxy-4-dedimethylaminotetracycline,4-dedimethylamino-7-chlorotetracycline,4-dedimethylamino-7-dedimethylaminotetracycline, the 2-nitrilo analogsof tetracycline, 4-dedimethylamino 12a-deoyotetracycline, tetracyclinesaltered at the 2-carbon position to produce a nitrile,4-dedimethylamino-7-chlorotetracycline,6-alpha-deoxy-5-hydroxy-4-dedimethylaminotetracycline,tetracyclonitrile, 6-alpha-benzylthiomethyltetracycline, the 2-nitriloanalog of tetracycline, 11-alpha-chlorotetracycline,7-chlortetracycline, 5 hydroxytetracycline,6-demethyl-7-chlortetracycline,6-demethyl-6-deoxy-5-hydroxy-6-methylenetetracycline,6-alpha-benzylthiomethylenetetracycline, a nitrile analog oftetracycline, a mono-N-alkylated amide of tetracycline,2-acetyl-8-hydroxy-1-tetracycline, 6-demethyl-6-deoxytetracycline,6-demethyl-6-deoxy-5-hydroxy-6-methylenetetracycline,2-acetyl-8-hydroxyl-1-tetracycline,4-hydroxy-4-dedimethylaminotetracycline,5a,6-anhydro-4-hydroxy-4-dedimethylaminotetracycline,6-demethyl-6-deoxy-4-dedimethylaminotetracycline,6-deoxy-8-demethyl-4-dedimethylaminotetracycline,6a-deoxy-5-hydroxy-4-dedimethylaminotetracycline, tetracyclines alteredat the 2-carbon position to produce a nitrile, pyrazole derivative oftetracycline, 7-chloro-6-demethyl-4-dedimethylaminotetracycline,11-apha-chlortetracycline, 4-dedimethylamino-7-chlortetracycline,4-de(dimethylamino)-tetracycline, 4-de(dimethylamino)-5-oxytetracycline,4-de(dimethylamino)-7-chlortetracycline,7-chloro-6-demethyl-4-dedimethylaminotetracycline,6-o-deoxy-5-hydroxy-4-dedimethylaminotetracycline,6-alpha-obenzylthiomethylenetetracycline,4-de(dimethylamino)-5-oxytetracycline,4-de(dimethylamino)-7-chlortetracycline,4-hydroxy-4-dedimethylaminotetracycline,6-alpha-deoxy-5-hydroxy-4-dedimethylaminotetracycline,4-de(dimethylamino)-tetracycline,4-de(dimethylamino)-7-chlortetracycline,7-chloro-6-demethyl-4-dedimethylaminotetracycline.dedimethylaminotetracycline 6-alpha-benzyl-thiomethylenetetracycline,11-alpha-chlortetracycline,6-demethyl-6-deoxy-5-hydroxy-6-methylenetetracycline,6-fluoro-demethyltetracycline, and the salts, conjugates, derivativesand combinations thereof.
 3. The method of claim 1, wherein said analogcomprises a form suitable for oral administion.
 4. The method of claim2, wherein said analog comprises a form suitable for topicalapplication.
 5. The method of claim 1, wherein said analog comprises aform suitable for administion by way of an injection or intravenousperfusion.
 6. The method of claim 1, wherein said analog comprises anamount suitable for providing a dosage of from about 0.1 mg/kg/day toabout 100 mg/kg/day.
 7. The method of claim 6, wherein said analogcomprises an amount suitable for providing a dosage of from about 10 mgper kg per day to about 50 mg per kg per day.
 8. The method of claim 7,wherein said analog comprises an amount suitable for providing a dosageof from about 20 mg per kg per day to about 25 mg per kg per day.
 9. Amethod for treating the biochemical formation of materials incidental toHIV or AIDS or which promote the advancement of HIV or AIDS or whichpromote the advancement of AIDS and other opportunistic diseases towhich a patient might become ill comprising administering to patients inneed of such treatment an anti-retroviral amount of non-antimicrobial,non-antibiotic, non-antibacterial chemically modified tetracycline (CMT)analogs.
 10. The method of claim 9 wherein the chemically modifiedtetracycline (CMT) is selected from the group consisting of: 4-dedimethylaliinotetracycline, 4 -dedimethylamino-oxytetracycline, 4-dedimethylamino- 7 -chlorotetracycline, 4 -hydroxy- 4-dedimethylaminotetracycline, 5 -alpha- 6 -anhydro- 4 -hydroxy- 4-dedimethylaminotetracycline, 6 -alpha-deoxy- 5 -hydroxy- 4-dedimethylaminotetracycline, 6 -dimethyl- 6 -deoxy- 4-dedimethylaminotetracycline, 4 -dedimethylamino- 11 -hydroxy- 12-alpha-deoxytetracycline, 12 -alpha-deoxy- 4deoxy- 4-dedimethylaminotetracycline, 6 -alpha-deoxy- 5 -hydroxy- 4-dedimethylaminotetracycline, 12 -alpha,4alpha-anhydro- 4-dedimethylaminotetracycline, minocycline-CMT, 7 -dimethylamino- 6-dimethyl- 6 -deoxy- 4 -dedimethylaminotetracycline, 6-alpha-benzylthiomethylenetetracycline, the 2 -nitrilo analogs oftetracycline (tetracyclinonitrile), the mono-N-alkylated wide oftetracycline, 6 -fluoro- 6 -demethyltetracycline,11alpha-chlorotetracycline, tetracycline pyrazole, 12-alpha-deoxytetracycline, 4 -dedimethylamino- 5 -oxytetracycline, 5-alpha, 6 -anhydro- 4 -hydroxy- 4 -dedimethylaminotetracycline, 12-alpha, 4alpha-anhydro- 4 -dedimethylaminotetracycline,tetracyclonitrile, 7 -chloro- 4 -dedimethylaminotetracycline, 12 -alpha-4 -deoxy- 4 -dedimethylaminotetracycline, 4 -dedimethylamino- 7-chlorotetracycline, 4 -dedimethylamino- 7 -dedimethylaminotetracycline,the 2 -nitrilo analogs of tetracycline, 4 -dedimethylamino-12alpha-deoxyotetracycline, tetracyclines altered at the 2 -carbonposition to produce a nitrile, 4 -dedimethylamino- 7-chlorotetracycline, 6 -alpha-deoxy- 5 -hydroxy- 4-dedimethylaminotetracycline, tetracyclonitrile, 6-alpha-benzylthiomethyltetracycline, the 2 -nitrilo analog oftetracycline, 11 -alpha-chlorotetracycline, 7 -chlortetracycline, 5-hydroxytetracycline, 6 -demethyl- 7 -chlorotetracycline, 6 -demethyl- 6-deoxy- 5 -hydroxy- 6 -methylenetetracycline, 6-alpha-benzylthiomethylenetetracycline, a nitrile analog oftetracycline, a mono-N-alkylated amide of tetracycline, 2 -acetyl- 8-hydroxy- 1 -tetracycline, 6 -demethyl- 6 -deoxytetracycline, 6-demethyl- 6 -deoxy- 5 -hydroxy- 6 -methylenetetracycline, 2 -acetyl- 8-hydroxyl- 1 -tetracycline, 4 -hydroxy- 4 -dedimethylaminotetracycline,5 -alpha, 6 -anhydro- 4 -hydroxy- 4 -dedimethylaminotetracycline, 6-demethyl- 6 -deoxy- 4 -dedimethylaminotetracycline, 6 -deoxy- 8-demethyl- 4 -dedimethylaminotetracycline, 6 -alpha-deoxy- 5 -hydroxy- 4-dedimethylaminotetracycline, tetracyclines altered at the 2 -carbonposition to produce a nitrile, pyrazole derivative of tetracycline, 7-chloro- 6 -demethyl- 4 -dedimethylaminotetracycline, 11-alpha-chlorotetracycline, 4 -dedimethylamino- 7 -chlorotetracycline, 4-de(dimethylamino)-tetracycline, 4 -de(dimethyl)- 5 -oxytetracycline, 4-de(dimethylamino)- 7 -chlorotetracycline, 7 -chloro- 6 -demethyl- 4-dedimethylaminotetracycline, 6 -alpha-deoxy- 5 -hydroxy- 4-dedimethylaminotetracycline, 6 -alpha-obenzylthiomethylenetetracycline,4 -de(dimethylamino)- 5 -oxytetracycline, 4 -de(dimethylamino)- 7-chlorotetracycline, 4 -hydroxy- 4 -dedimethylaminotetracycline, 6-alpha-deoxy- 5 -hydroxy- 4 -dedimethylaminotetracycline, 4-de(dimethylamino)-tetracycline, 4 -de(dimethylamino)- 7-cholorotetracycline, 7 -chloro- 6 -demethyl- 4-dedimethylaminotetracycline, dedimethylaminotetracycline, 6-alpha-benzyl-thiomethylenetetracycline, 11 -alpha-chlorotetracycline, 6-demethyl- 6 -deoxy- 5 -hydroxy- 6 -methylenetetracycline, 6-fluoro-demethyltetracycline, and the salts, conjugates, derivatives andcombinations thereof.
 11. The method of claim 9 wherein said analogcomprises a form suitable for oral administration.
 12. The method ofclaim 10 wherein said analog comprises a form suitable for topicalapplication.
 13. The method of claim 9 wherein said analog comprises aform suitable for administration by way of an injection or intravenousperfusion.
 14. The method of claim 9 wherein said analog comprises anamount suitable for providing a dosage of from about 0.1 mg/kg/day toabout 100 mg/kg/day.
 15. The method of claim 14 wherein said analogcomprises an amount suitable for providing a dosage of from about 10 mgper kg per day to about 50 mg per kg per day.
 16. The method of claim 15wherein said analog comprises an amount suitable for providing a dosageof from about 20 mg per kg per day to about 25 mg per kg per day.